Free radical scavenging activities of some indigenous plants of Bangladesh Mohammad S. Rahman’, Mohammed Z. Rahman, 1S. Ahmed and Mohammad A. Rashid" ‘Department of Pharmacy, The University of Asia Pacific, Dhaks-1208, Bangladesh, Department of Pharmaceutical Chemisty, Faculty of Pharmacy, Unversity of Dhaka, Dhaka-1000, Bangladesh Centre for Biomedical Research, University of Dhaka, Dhak-1000, Bangladesh “Directorate of Drug Administration, Motel, Dhaka Abstract: Five indigenous plants, Bougainulea glabra, Gmelina hystrix, Jatropha pandunfoia, Pereskiagrandiflia and Xyiocarpus moluecenss, have been investigated for their antioxidant activity. The extractives were subjected to assay by 1,1-dipheny-2-pienhydrazyl (DPPH) for evaluation of free radial scavenging property. The ICx of the organic extracts ranged from 323 to 22 g/l. The crude methanol extract of X-moluccensis demonstrated potent fre radical scavenging activity JC» 2 g/ml) as compared to {ert-butyl-t-hydroxjtoluene (standard) that showed an IOs value of 81Sug/mi The results primarily suggest the presence of potent oxidation inhibitory principles in the bark of X moluccensis. Keywords: 1,1-Dipheny-2-pleryl hydrazyl, antioxidant; fee radical scavenger Introduction There has been a worldwide positive move towards the use of traditional medicines due to the concern over the more invasive, expensive and potentially toxic mainstream modern practices (Harnett ef al, 2005 WHO, 2002). ts popularity is due to the desire for more personalized health care and greater public access to health information (Greene and Peterin, 2002), Free radicals play a erucial role in the development of tissue damage in various hnuman diseases such as cancer, aging, neurodegenerative disease, arteriosclerosis and pathological events in. living organisms (Erdemogli et al, 2008; Gutteridge, 1994). Antioxidants may have an important role in the prevention of these diseases. There is an increasing interest in the antioxidant effects of compounds derived {rom plants, which could be relevant in relation to their nutritional incidence and their role in health and disease (Takahashi et a, 1982; watsuki ot af, 1988; Steinmetz ‘and Potter, 1996; Wang et al, 1999; Aruoma, 1998; Bandoniene et af, 2000; Pieroni et al, 2002; Couladis et al, 2003). A number of reports on the isolation and testing of plant derived antioxidants have been described during the past decade (Shahidi et af, 1992; Volioglu et al, 1998; Pietta et al, 1998). Extensive literature survey has revealed that no research has been ‘conducted on the antioxidant activities of the selected “Autir fr Corspondence plants (htipy/www.nebinim.nih.gov/pubmed). AS a part of our effort directed to the search for such molecules from natural sources, we studied the methanol extracts of &. glabra, G. hystrix, J. pandurifolia, .grandifolia and X. moluccensis, and we, herein, report the results of our preliminary investigation Materials and methods Plant Materials: Leaves of Bougainvilea glabra (Family: Nyctaginaceae), Gmelina hystrix (Family Lamiaceae alt. Labiatae), Jatropha _pandurifolia (Family: Euphorbiaceae), Pereskia grandifolia (Family Cactaceae) and Xylocarpus moluccensis (Family: Meliacese), were collected from Dhaka in the month of January 2007 and voucher specimens have been deposited in Bangladesh National Herbarium (BNH). The leaves the plants were first separated from the plant, cut into small pieces and air-dried for several days. The pieces were then oven dried for 24 hours at considerably low temperature to facilitate size reduction through grinding Extraction: The air-dried and powdered leaves of the plants were separately extracted with methanol for 15 days at room temperature with occasional shaking and stirring. It was then filtered through a fresh cotton Bangladesh Pharmacoutical Joural, Vol. 19, No 1, January 2010 68 ISSN no,’ 0501-8608plug and finally with a Whatman No.1 filter paper The volume of the filtrate was then reduced using a Buchii Rotavapor at low temperature and pressure Subsequent evaporation of solvents afforded extracts of B, glabra (72 9), J. pandurifolia (6.6 g), G. hystrix (08 9), P. grandifolia (0.78 g) and X ‘moluccensis (0,84 g), Solvent-solvent partitioning of B. glabra and J. pandurifolia was done using the protocol designed by Kupchan and modified by Van Wagenen et al. (1993) to provide mhexane, carbon tetlachloride and chloroform soluble fractions. The crude methanol extract of G. hystrix P. grandifolia and X moluccensis were not subjected to partitioning due to small amount of sample. Antioxidant activity: The antioxidant activity (free radical scavenging activity) of the extracts on the stable radical 1-diphenyl-2-pierylhydrazyl (DPPH) was. determined by the method developed by Brand-Williams et al, 1995. In the experiment, 2.0, mg of each of the extracts was dissolved in ‘methanol, Solution of varying concentrations such a8 500-, 250-, 125-, 62.50-, 31.25-, 15.62-, 78125- 3.91-, 1.95 and 0.98 ig/ml were obtained by serial dilution technique. An aliquot of 2 ml of the methanol solution of the extract of each concentration was mixed with & ml of a DPPH-methanol solution (20 mg/L) and allowed to stand for 20 minutes for the reaction to occur. Then the absorbance was determined at 517 nm and from these values corresponding percentage of inhibitions were calculated by using the following equation: ‘inhibition = [1- (ABSwaph / ABSe 9) x10 Finally, the 9% inhibitions were plotted against respective concentrations used and from the graph ICs was calculated. Here, —_tert-butyl-1 -hydroxytoluene (BHT), @ potential antioxidant, was used as the positive control. Statistics: Al the analysis was carried out in triplicate and the results are expressed as mean + SD. Results and discussion The extractives of B. glabra, G. hystrix, pandurifolia, free radical scavenging activity and results are presented in Table-1. The antioxidants act either by scavenging various types of free radicals derived {rom oxidative processes, by preventing free radical formation through reduction precursors or by chelating metals (Burton and Ingold, 1984; Bors et al, 1984; Mark e¢ al, 1994). The reduction of DPPH assay has been used to detect products with antioxidant activity as free radical scavengers Clubaro et al, 1996; Cavin et al, 1998; Gamez et al 1998). In this study, all the extractives were shown to possess significant DPPH radical scavenging activity. X moluccensis was found to have the highest antioxidant activity with an ICse value of 22 ig/ml. However, significant antioxidant activity was noticed by the chloroform soluble fraction of methanolic extract of J. pandurifoia VCs 91 jg/m), carbon tetrachloride soluble fraction of methanolic extract of B. glabra (ICs 87 ug/ml), hexane soluble fraction of methanolic extract of B. glabra (ICx 108 lig/mD and carbon tetrachloride soluble fraction of methanolic extract of J pandurifolia(ICs» 104 wa/m)). On the other hand, moderate antioxidant activity was revealed by methanolic extract of leaves of J. pandurifolia (Css 160 g/m), and its n-hexane soluble fraction (ICs 165 ug/ml) and methanolic extract of leaves of G. hystrix (Css 170 yg/mD. The methanol extracts of B. glabra and P. grandifolia demonstrated weak free radical scavenging activity with ICs of 323 ig/ml and 450 ug/ml, respectively Therefore, it can be concluded that all the plants have great potential to act as antioxidant, which also indicates the presence of secondary metabolites having antioxidant activities. These plants could be subjected for extensive chromatographic separation ‘and putification processes to isolate the bioactive compounds for the discovery of leads for further development. P. grandifola and X. moluccensis, were assessed (or frma=——paumiraezin Bangladesh Pharmacoutical Joural, Vol. 19, No 1, January 2010 69 ISSN no,’ 0501-8608References Aruoma, O11, (1888). 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